Mice are the most commonly used preclinical animal model, but protein analytics of murine cerebrospinal fluid (CSF) remains challenging because of low CSF volume (often <10 µl) and frequent blood contaminations. We developed an improved CSF sampling method that allows routine collection of increased volumes (20-30 µl) of pure CSF from individual mice, enabling multiple protein analytical assays from a single sample. Here, we have analysed the CSF of 3, 6, and 12 months old wild-type mice. Additionally, we collected murine CSF of the same mice at 3 and 6 months of age to compare repeated and single CSF sampling. Furthermore, we analysed human CSF samples as a reference.